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J. Biol. Chem., Vol. 278, Issue 20, 17918-17926, May 16, 2003
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From the Department Biologie I, Bereich Genetik,
Ludwig-Maximilians-Universität München, Maria-Ward-Strasse
1a, D-80638 Munich, Germany
The molecular basis of constitutive gene
activation is largely unknown. The yeast profilin gene
(PFY1), encoding a housekeeping component of the actin
cytoskeleton, is constitutively transcribed at a moderate level. The
PFY1 promoter dispenses with classical transactivators and
a consensus TATA box; however, it contains a canonic site for the
abundant multifunctional nuclear factor rDNA enhancer-binding protein
(Reb1p) combined with a dA·dT element. Reb1p binds specifically
in vitro. Mutation of this site reduces PFY1
expression to about 35%. A nucleosome-free gap of about 190 bp is
centered at the genomic Reb1p binding site in vivo and
spans the presumptive core promoter and transcriptional initiation
sites. Nucleosomes at the border of the gap are positioned. Mutation of
the Reb1p motif in the genomic PFY1 promoter abolishes
nucleosome positioning, fills the gap with a non-positioned nucleosome,
and reduces transcription by a factor of 3. From permutation studies we
conclude that Reb1p induces a strong bend into the DNA. Phasing analyses indicate that it is directed toward the major groove. The data
suggest that Reb1p plays an architectural role on DNA and that
Reb1p-dependent DNA bending leads to a DNA conformation that is incompatible with packaging into nucleosomes and concomitantly facilitates constitutive transcription. In the absence of other transcription activators, Reb1p excludes nucleosomes and moderately stimulates transcription by distorting DNA.
Reb1p-dependent DNA Bending Effects Nucleosome
Positioning and Constitutive Transcription at the Yeast Profilin
Promoter*
,
*
This work was supported by Deutsche Forschungsgemeinschaft,
Sonderforschungsbereich Grant 190-TP B6 (to W. B.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.:
49-89-2180-6176; Fax: 49-89-2180-6160; E-mail:
M.Angermayr@lrz.uni-muenchen.de.
§
Present address: Lichtenstein Pharmaceutica GmbH und Co.,
Industriestrasse 10, D-82256 Fürstenfeldbruck, Germany.
Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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